Immunocytochemistry/Immunofluorescence: Apolipoprotein E/ApoE Antibody (WUE-4) [ABIN2587851] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ApoE (WUE-4) [ABIN2587851] at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Application Details

Application Details

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Application Notes

Western Blot 2 μg/mL, Flow Cytometry 1 μg per million cells, ELISA 1:100-1:2000, Immunohistochemistry 1:50-1:200, Immunocytochemistry/Immunofluorescence 1:200, Immunoprecipitation 1:10-1:500, CyTOF-readyThis ApoE antibody is useful for Western blot, ELISA, Immunohistochemistry and Immunoprecipitation. In Western blot a band is observed at ~36 kDa, representing the ApoE protein. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 26564908)

Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol

Protocol specific for ApoE Antibody Western Blot Protocol1. Perform SDS-PAGE (4-12 % MOPS) on samples to be analyzed, loading 40ug of total protein per lane.. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus. . Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.. Rinse the blot in TBS for approximately 5 minutes.. Block the membrane using 5 % non-fat dry milk + 1 % BSA in TBS, 1 hour at room temperature.. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.. Dilute the mouse anti-ApoE primary antibody in blocking buffer and incubate 1 hour at room temperature.. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.. Apply the diluted mouse-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer'sinstructions) and incubate 1 hour at room temperature.. Wash the blot in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.

Immunocytochemistry/Immunofluorescence: Apolipoprotein E/ApoE Antibody (WUE-4) [ABIN2587851] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ApoE (WUE-4) [ABIN2587851] at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.